KR20110081327A - Treatment of t-cell lymphoma using 10-propargyl-10-deazaaminopterin - Google Patents

Abstract

T cell lymphoma is treated by administering to a patient suffering from T cell lymphoma a therapeutically effective amount of 10-propargyl-10-deazaaminopterin. Remission is observed in human patients, even with drug resistant T cell lymphoma at weekly dosages levels as low as 30 mg/m2. In general, the 10-propargyl-10-deazaaminopterin is administered in an amount of from 30 to 275 mg/m2 per dose.

Description

Treatment of T-cell lymphoma using 10-propargyl-10-deazaaminopterin}

This application is filed under US Regular Application No. 30 May 2004. 60 / 521,593 is claimed as a priority and the regular application is incorporated herein by reference.

According to the present invention, T cell Non-Hodgkin's Lymphoma is treated using 10-Propargyl-10-Deazaaminopterin (PDX). Thus, according to one aspect of the present invention, there is provided a method for the treatment of T cell non-Hodgkin's lymphoma, which comprises administering a therapeutically effective amount of PDX to patients suffering from lymphoma. Preliminary clinical results in humans show that these treatments are particularly effective, even for lymphomas that have been refractory to other treatment modalities.

This application relates to the treatment of T-cell lymphomas and to the use of the 10-propargyl-10-deazaaminopterin combination.

10-Propargyl-10-Deazaaminopterin ("PDX" or "10-propargyl-10dAM") has been tested in the treatment of carcinomas and is useful in some cases. It is one of many compounds that are found to be. This compound having the structure shown in Figure 1 is "synthetic and anticancer activity of 10-propargyl-10-deazaaminopterin" by DeGraw et al. ( J. Medical Chem . 36: 2228-2231 (1993)) and act as growth inhibitors in the murine animal L1210 cell line and weakly act on the enzyme dihydrofolate reductase ("DHFR"). Showed. In addition, when using a breast cancer model of E0771 murine, any results for the anticancer characteristics of the compounds are shown. Due to the small number of rats used in the experiment (three per dose), the absence of any standard deviation information to determine the reliability of the results, and the fact that the highest dose used was actually toxic to the rats, these results were not clear. Was not. Nevertheless, assuming that these results have certain expectations for the efficacy of the drug in treating human cancer, the same values of resistance are probably slightly better or comparable to methods of methodotrexate. It seems to be optimal to anticipate drugs that have.

PCT Publication No. WO98 / 02163 is far superior to Mesotrexate ("MTX") when PDX compositions purified to higher levels for human cancer efficacy have been tested in xenograft models, and even more recently clinical candidates. It is a surprising discovery that it is superior to edatrexate ("ETX"). In addition, 10-propargyl-10dAM has shown an amazing ability to treat cancer so that there is no evidence that the cancer has grown for several weeks after discontinuation of treatment. Thus, highly purified compositions comprising 10-propargyl-10dAM can be used in accordance with the present invention for treating cancer, including both solid and leukemia. The composition is described for use in the treatment of human breast cancer and human lung cancer.

Subsequent studies on PDX have shown to be useful both as PDX itself and in combination with other therapeutic agents. For example, Sirotnak et al. (Clinical Cancer Research Vol. 6, 3705-3712 (2000)) is a cMOAT / MRP inhibitors of probenecid coadministration of (probenecid) and PDX, such as the plasma membrane ATP in kinase is It has been reported to greatly increase the efficacy of PDX on human solid cancer in vivo . PDX, and combinations of platinum and PDX based on chemotherapy drugs, have been shown to be effective against mesothelioma (Khokar et al., Clin. Cancer Res. 7: 3199-3205 (2001)).

The term "lymphomas" refers to Non-Hodgkins Lymphoma (NHL); Diffuse large B-cell lymphoma (DLBCL); Follicular lymphoma (FL); Hodgkin's Disease; Burkitt's Lymphoma; Cutaneous T cell lymphoma; It indicates a number of disease states, including primary central nervous system lymphoma and lymphoma metastases. In most cases, lymphomas are characterized by the presence of B cells with cancer. However, in T cell lymphoma, such disease state is characterized by T lymphocytes with cancer.

This application relates to the use of 10-propargyl-10-deazaaminopterin in the treatment of T-cell lymphoma.

T-cell lymphoma is a lymphoma in which a patient's T cells turn out to be cancerous. T cell lymphoma includes a variety of conditions without the following limitations.

(a) Lymphoblastic lymphomas with malignancy in the primordial lymphoid progenitors from the thymus;

(b) T cell prolymphocytic leukemia, T-cell granular lymphocytic leukemia, aggressive NK-cell leukemia, cutaneous T cell lymphoma (Normal carcinosarcoma / Terrisne syndrome), anaplastic large cell lymphoma, T cell type, enteropathy-type T cell lymphoma, HTLV-1 Mature or peripheral T cell tumors, including adult T cell leukemia / lymphoma, angioimmunoblastic T cell lymphoma, and subcutaneous panniculitic T cell lymphoma

(c) Peripheral T cell lymphoma, which initially includes a lymph node thyroid-dependent layer but never grows in a typical follicular form.

In one embodiment of the invention, the composition consists of "very purified" PDX. As used in the specification and claims of the present invention, a "very purified" composition consists of 10-deazaaminopterin, which can substantially interfere with the anticancer activity of other folic acid derivatives, specifically PDX. Contains no PDX. Within the scope of the present invention, the composition may comprise additional, non-folic acid therapeutic agents, as well as delivery or excipients for formulating the PDX in the form of a suitable dosage unit for therapeutic use.

PDX is the DeGraw paper or US Patent No. 5,353,751, which can be synthesized using the method disclosed in Example 7, which patent is incorporated herein by reference. HPLC evaluation of the product produced by this method shows that there is a significant amount (˜4.6%) of impurity A having a retention time consistent with 10-deazaaminopterin (Figure 2). Therefore, if this synthetic approach is applied, further purification is needed beyond that described in DeGraw et al. Such purification can be performed by additional HPLC or crystallization to remove 10-deazaaminopterin and other folic acid derivatives that may be present.

3 shows the HPLC of a highly purified preparation consisting essentially of 10-propargyl-10dAM according to the present invention made using the method disclosed in Example 1. FIG. In this case, the amount of PDX (as determined by HPLC peak area) is close to 98%, and although there is a small change in baseline in this area, the peak corresponding to 10-deazaaminopterin is It was not detected by the advanced software.

PDX was used in the Phase I / II study for patients who were pathologically infected with aggressive lymphoma, including three patients with drug-resistant T cell lymphoma. A summary of the object is obtained below. In addition, each of these patients was treated with folic acid (1 mg / m ² daily and 1 weekly Bmg (1 mg / m ² once a month) supplements prior to PDX treatment.

Patient 1.

Diagnosis: Peripheral T Cell Lymphoma, Stage IV

Population: 48-year-old male

Previous treatment: CHOP 4 cycles (July 2002-November 2002)-incurable

           2 rounds of ICE (December 2002)-Nanchi

           Campath (March 2003-June 2003)-Mixed reactions

Premorbid staging: a wide range of diseases of skin diseases

Treatment for the study: PDX 135 mg / m ² single dose

Toxicity: Grade 3 stomatitis; Grade 3 neutropenia; blood poisoning

Reaction: Intrinsically alleviated according to PET scan

Opinion: Ultimately, the patient died after onset of bacteremia and sepsis from exposed skin lesions on Gram-positive bacteria.

Patient 2.

Diagnosis: Lymphoblastic Lymphoma, Precursor T-Cells, Stage IV

Population Trends: Women 65

Previous Treatment: Chemotherapy of L20-complex combination since May 2002, administered for 2 years or more. Received MTX from May 2002 to February 2004. Disease recurred in December 2004.

Pretreatment Arm: Extensive Recurrence

Treatment for the study: PDX 30 mg / m ² every 3 weeks

                  Completed in three cycles

Toxicity: None

Reaction: fully relaxed according to PET and CT scan

Opinion: Patients with intrinsic mesotrexate-resistant diseases and massive sinus based on disease began to heal after a single dose of PDX.

Patient 3.

Diagnosis: T-Cell Lymphoma Associated with HTLV

Population trend: 38 year old male

Previous Treatment: Combination Chemotherapy of EPOCH-Injection

           From October 2003 to February 2004

Pretreatment Arm: Onset on Left Armpit

Treatment for the study: PDX 30 mg / m ² per week, 3 times every 4 weeks, 2 cycles

Toxicity: None

Reaction: fully relieve

Opinion: Clinically obvious disease disappears completely at the end of the first cycle, very well tolerated, no toxicity

Patient 4:

Diagnosis: Fatty Cell T-Cell Lymphoma

Population trend: 25 year old male

Previous treatment: Ontak (intractable); 9 / 02-11 / 02; Targretin and IFN α1 / 03-10 / 03 (resistant and partially mitigated); CHOP 4 / 04-6 / 04; ICE 6/04, CyPen 7 / 04-8 / 04, Targretin / MTX 9/04 from 2/05

Treatment for Research: PDX 30mg / m ² 4 times per week

Response: Completely clinically mitigated according to PET

Toxicity: None

Comments: Treating subcutaneous lesions, too many, countless ulcer granular lesions

To date, only four patients with T-cell lymphoma have been treated with PDX, all four patients have met the criteria for complete relief, and even met the criteria for complete relief even on the basis of sensitive PET imaging techniques. It was. Interestingly, patients treated at 135 mg / m ² received only one dose of medication and had a dramatic response to treatment, while the remaining patients received only small amounts of excised doses according to the weekly schedule.

For use according to the invention, PDX is conveniently formulated as part of a pharmaceutical preparation. The particular dosage form will vary depending on the method of administration, but may also include tablets, capsules, oral solutions, infusion solutions for intravenous, intramuscular, or intraperitoneal administration. One suitable dosing schedule includes administering 150 mg / m ² every two weeks. Of course, depending on the tolerance of the individual patient, or if more frequent administration is adopted, it may be indicated to be used at a lower level. For example, a level similar to 40-120 mg / m ² is suitable for body surface area / day. 3 shares the capacity of the week and stop in a while, and administered at a dose of 30mg / m ² weeks, weekly and administered for 6 weeks at a dose of 30mg / m ² and stopped for a week, or gradually week PDX six-week planned for It is also appropriate to increase. If a lower frequency of administration should be used, higher levels may be used. Thus, in a general sense, dosages of 30-275mg / m ² in accordance with various schedules of administration, for example, the amount of bi-weekly administration of 135 ~ 275mg / m ^2, may suitably be used so that the 1-week dose 30 ~ 150mg / m ² the amount of have. U.S. Patent No. incorporated herein by reference Using an experimental design similar to that disclosed in 6,323,205, it is within the skill of one in the art to determine the appropriate dosage.

PDX includes vinca alkaloids such as vinblastine, navelbine and vindesine; Nucleic acid analogues such as probenicid, gemcitabine, 5-fluorouracil and cytarabine; Alkylating agents such as cyclophosphamide or ifosfamide; Cisplatin or carboplatin; Leucovorin; Taxanes such as paclitaxel or doketaxel; Anti-CD20 monoclonal antibodies with or without radioisotopes; And other cytotoxic or anticancer compounds, including antibiotics such as doxorubicin and mitomycin. PDX may also be used in combination with several of these other anticancer agents or in combination with growth factor inhibitors and anti-angiogenic agents.

PDX and other drugs may be administered simultaneously or may be used in combination as part of a conventional treatment regimen, where PDX and other drugs are administered at different times. For example, other medications may be administered before, immediately after, or after a period of time (eg, 24 hours) for PDX administration. Thus, for this purpose of application, the term administration generally refers to the continuous administration of drugs in either order in the simultaneous treatment of drugs or in the treatment regimen between the drugs separately in time or in time, unless otherwise specified. It shows administration.

To reduce the side effects of therapy, PDX is suitably used in combination with folic acid and vitamin B12 supplements. For example, patients are treated with folic acid (1 mg / m ² daily and 1 week / week (1 mg / m ² once a month) prior to starting PDX treatment.

Figure 1 shows the structure of PDX and method trexate.
2 shows the HPLC of a non-pure 10-propargyl-10dAM preparation prepared according to the prior art.
3 shows the HPLC of a highly purified PDX preparation according to the present invention.
4 shows a synthetic schematic diagram useful in preparing compounds according to the invention.

Example 1

4 shows a synthetic schematic that is useful when preparing 10-propargyl-10-dAM according to the present invention. To a mixture of 60% NaH (1.06 g, 26.5 mmol) in an oil dispersion was cooled to 0 ° C. in 18 ml of THF dehydrated with sieves. The cooled mixture was treated with a solution of homoterephthalic acid dimethyl ester (5.0 g, 24 mmol, compound 1 in Figure 4) in dried THF (7 ml) and the mixture was stirred at 0 ° C. for 1 hour. Propargyl bromide (26.4 mmol) was added and the mixture was stirred for additional 1 h at 0 ° C. and then for 16 h at room temperature. The resulting mixture was treated with 2.4 mL of 50% acetic acid and then poured into 240 mL of water. The mixture was extracted with ether (150 mL twice). Ether extracts were combined, dried over Na ₂ S0 ₄ and concentrated to orange-yellow oil. Silica gel (230-400 mesh, 600 ml) chromatography with an eluent of cyclohexane-EtOAc (8: 1) chromatographed product α-propargyl homoterephthalic acid uniformly seen in TLC (cyclohexane-EtOAc, 3: 1) Dimethyl ester (Compound 2) was obtained as a white solid (4.66). However, mass spectral analysis of this product showed that it was a mixture of the desired compound 2 and the dipropargylated compound. Starting material 1 was not detected. HPLC showed that the ratio of mono-propargylated product to di-propargylated compound was about 3: 1. The dipropargylated product, unlike compound 1 , could not produce unwanted byproducts in the next step of the reaction, which was suitable for conversion to compound 3. The absence of starting material 1 in the product to proceed with synthesis is very important to avoid the continuous production of 10-dAM during the conversion process leading to the final product, because complete removal from 10-propargyl-10-dAM Because it is very difficult.

0.36 g (9 mmol) of 60% NaH in an oil dispersion was added to 10 mL of dehydrated THF to form a mixture, and the mixture was cooled to 0-5 ° C. The cooled mixture was treated dropwise with a solution of the first reaction product (Compound 2) (2.94 g, 12 mmol) in 10 mL of dehydrated DMF and then stirred at 0 ° C. for 30 minutes. After cooling to -25 [deg.] C., 2,4, diamino-6- (bromomethyl) -pteridine hydrobromide-0.2.2.-propanol (1.00 g, 2.9 mmol) in 10 mL of dehydrated DMF. Was added dropwise while maintaining the temperature near -25 ° C. The temperature of the stirred mixture was allowed to rise to -10 ° C over a period of 2 hours. After an additional 2 hours at −10 ° C., the temperature was allowed to rise to 20 ° C .; Stirring was continued for longer than 2 hours at room temperature. The reaction was then adjusted to pH 7 by addition of solid CO ₂ . After concentration in vacuo to remove solvent, the residue was stirred with diethyl ether, insoluble material in ether was collected, washed with water and water removed in vacuo to yield 1.49 g of crude product. The crude product was dissolved in CHCl ₃ -MeOH (10: 1) to apply a silica gel column. Elution with the same solvent system showed homogeneity in TLC and methyl 10-propargyl-10-carbomethoxy-4-deoxy-4-amino-10-deazaphteroic acid at 40% yield Ester (compound 3) was obtained (485 mg).

A stirred suspension of compound 3 (400 mg, 0.95 mmol) in 2-methoxyethanol (5 mL) was treated with water (5 mL) and then with 10% sodium hydroxide solution (3.9 mL). The mixture was stirred at room temperature for 4 hours, during which time the solution appeared. The solution was adjusted to pH 8 with acetic acid and concentrated under high pressure vacuum. The resulting residue was dissolved in 15 mL of water and acidified to pH 5.5-5.8 to form a precipitate. The precipitate was collected, washed with water and dried in vacuo to recover 340 mg of compound 4 (91% yield). HPLC analysis indicated that the purity of the product was 90%.

Compound 4 (330 mg) was placed in 15 mL of DMSO and heated at 115-120 ° C. for 10 minutes to remove carboxyl groups. After 10 minutes analysis by HPLC confirmed that the conversion was essentially complete. DMSO was removed by distillation in vacuo (bath temperature 40 ° C.). The residue was stirred with 0.5 NaOH to give a clear solution. Acidification with 1N HCl to pH 5.0 afforded 10-propargyl-4-deoxy-4-amino-10-deazaphteroic acid (Compound 5) as a yellow solid, 70% yield. HPLC indicated the purity of the product at 90% at this stage.

BOP (benzotriazol-1-yloxytris (dimethylamino) phosphonium hexafluorophosphate) in DMF (10 mL) containing triethylamine (148 mg, 1.46 mmol) (287 mg, 0.65 mmol, Compound 5 (225 mg, 0.65 mmol) was reacted with dimethyl L-glutamate hydrochloride (137 mg, 0.65 mmol) using an Aldrich Chemical) reaction. The mixture was stirred at 20-25 ° C. for 3 hours and then distilled to remove moisture. The residue was stirred with water, the crude product insoluble in water was collected and the water was removed in vacuo. The crude product (350 mg) was purified by silica gel chromatography with eluent of CHCl ₃ -MeOH (10: 1) containing triethylamine (0.25% by volume) to give TLC (CHCl ₃ -MeOH, 5: In 1), 165 mg (compound 6, 50% yield) of 10-propargyl-10-deazaaminopterin dimethyl ester was recovered.

Compound 6 (165, 0.326 mmol) was suspended in 10 mL of stirred MeOH with 0.72 mL (0.72 meq) of 1N NaOH. Stirring was continued at room temperature and brought to a solution state after a few hours. The solution was kept at 20-25 ° C. for 8 hours and then diluted with 10 ml of water. Methanol was removed by evaporation under reduced pressure, and the concentrated aqueous solution was kept at 20-25 ° C. for 24 hours. Then, HPLC showed that ester hydrolysis took place completely. The clear aqueous solution was acidified with acetic acid to pH 4.0 to precipitate 10-propargyl-10-deazaaminopterin as a pale yellow solid. The collected, washed with water and dehydrated in vacuo weighed 122 mg (79% yield). Elemental analysis, analysis by hydrogen NMR and mass spectroscopy were entirely consistent with the designated structures. HPLC analysis indicated 98% purity and confirmed that the product was free of 10-deazaaminopterin.

Claims (20)

A pharmaceutical composition for the treatment of T cell lymphoma, comprising a pharmaceutically acceptable base addition salt of 10-propargyl-10-deazaaminopterin. The pharmaceutical composition of claim 1, wherein the salt of 10-propargyl-10-deazaaminopterin is substantially free of 10-deazaaminopterin. The pharmaceutical composition of claim 1, wherein the base addition salt is a sodium salt. The pharmaceutical composition of claim 1, wherein the pharmaceutical composition is used to administer a salt of 10-propargyl-10-deazaaminopterin in an amount of 30 to 275 mg / m ² per dose. A pharmaceutical composition that is formulated. The pharmaceutical composition of claim 1, wherein the T cell lymphoma is peripheral T cell lymphoma. The pharmaceutical composition of claim 1, wherein the T cell lymphoma is a relapsed or refractory peripheral T cell lymphoma. The pharmaceutical composition of claim 5 or 6, wherein the T cell lymphoma is peripheral T cell lymphoma excluding mycoses fungoides. 8. The method of claim 7, wherein the peripheral T cell lymphoma is T cell prolymphocytic leukemia; T-cell granular lymphocytic leukemia; Aggressive NK-cell leukemia; Cutaneous T-cell lymphoma, except fungal sarcoma; Anaplastic large cell lymphoma (T cell type); Enteropathy-type T cell lymphoma; Adult T-cell leukemia / lymphoma; Angioimmunoblastic T cell lymphoma; Subcutaneous panniculitic T cell lymphoma; And a peripheral T cell lymphoma, initially comprising a lymph node thyroid-dependent layer, but never progressing in a typical follicular form. The pharmaceutical composition of claim 8, wherein the peripheral T cell lymphoma is subcutaneous fatty stratitis T-cell lymphoma. Use according to claim 8, wherein the peripheral T cell lymphoma is Sezary syndrome. The pharmaceutical composition of claim 8, wherein the adult T-cell leukemia / lymphoma is human T-lymphotropic virus-1 (HTLV-1) associated T-cell lymphoma / leukemia . 4. The pharmaceutical composition of claim 1, wherein the pharmaceutical composition provides an amount of 30 to 275 mg / m ² per administration of a salt of 10-propargyl-10-deazaaminopterin. A pharmaceutical composition that is formulated for. The pharmaceutical composition according to any one of claims 1 to 3, wherein the pharmaceutical composition is formulated for weekly administration of a salt of 10-propargyl-10-deazaaminopterin. The pharmaceutical composition of claim 13, wherein the pharmaceutical composition is formulated to administer a salt of 10-propargyl-10-deazaaminopterin in an amount of 30 mg / m ² per dose. . The pharmaceutical composition of claim 13, wherein the pharmaceutical composition is formulated to administer a salt of 10-propargyl-10-deazaaminopterin in an amount of 30 to 150 mg / m ² per dose. Pharmaceutical composition. The pharmaceutical composition according to any one of claims 1 to 3, wherein the pharmaceutical composition is formulated for biweekly administration of a salt of 10-propargyl-10-deazaaminopterin. The pharmaceutical composition of claim 16, wherein the pharmaceutical composition is formulated to administer a salt of 10-propargyl-10-deazaaminopterin in an amount of 135 to 275 mg / m ² per dose. Pharmaceutical composition. 4. The pharmaceutical composition of claim 1, wherein the pharmaceutical composition is formulated for administering a salt of 10-propargyl-10-deazaaminopterin in one or more 7 week cycles. Wherein each cycle is administered once a week for 6 weeks at a content of 30 to 150 mg / m ² per dose and then stopped for 1 week. The pharmaceutical composition of claim 18, wherein the pharmaceutical composition is formulated to administer a salt of 10-propargyl-10-deazaaminopterin in an amount of 30 mg / m ² per dose. . 4. The pharmaceutical composition of claim 1, wherein the pharmaceutical composition further comprises folic acid and vitamin B ₁₂ , wherein the pharmaceutical composition comprises a salt of 10-propargyl-10-deazaaminopterin. A pharmaceutical composition formulated for separate, sequential or simultaneous administration of folic acid and vitamin B ₁₂ .

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